Letalidad de citocalasina B y otros compuestos aislados del hongo Aspergillus spp. (Trichocomaceae) endófito de Bauhinia guianensis (Fabaceae) / Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae)

Los hongos endofíticos son hongos que colonizan los tejidos internos de las plantas; varios compuestos biológicamente activos se han aislado a partir de estos hongos. Existen pocos estudios de compuestos aislados de hongos endófitos de plantas amazónicas. Por lo tanto, este estudio tuvo como objetivo el aislamiento y la identificación estructural de ergosterol (1), peróxido de ergosterol (2), mevalonolactona (3), citocalasina B (4) y citocalasina H (5) a partir de Aspergillus spp. EJC 04, un hongo endofítico de Bauhinia guianensis. La citocalasina B (4) y el derivado diacetato de citocalasina B (4a) mostraron una alta letalidad en el ensayo de Artemia salina. Esta es la primera aparición de citocalasinas en hongos endófitos amazónica de B. guianensis

Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis

Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae).

Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis.

Immobilized proteoliposome affinity chromatography for quantitative analysis of specific interactions between solutes and membrane proteins. Interaction of cytochalasin B and D-glucose with the glucose transporter Glut1.

An affinity gel bed was prepared by reconstitution of a transmembrane protein, the human red cell glucose transporter (Glut1), followed by steric immobilization of the proteoliposomes in small and rigid gel beads by freeze-thawing. The specific interactions between the reconstituted Glut1, the transport inhibitor cytochalasin B (CB), and the transported solute D-glucose were analyzed by isocratic chromatography of CB on the Glut1-proteoliposome gel bed. Specific retardation of CB which decreased upon inclusion of the competitor D-glucose in the eluent was observed on-line. The equilibrium constants for CB and D-glucose interaction with Glut1 (Kd 1.5 x 10(-7) M and 67 mM, respectively) obtained by use of equations derived for the affinity chromatographic analysis were consistent with values obtained by others by conventional methods. Effects of liposome composition, pH, and time on the CB binding activity of Glut1 were studied. Reconstitution of a membrane protein into a lipid environment and steric immobilization of the proteoliposomes favor retention of the protein activity. Immobilized proteoliposome affinity chromatography (IPAC) is a novel, powerful method for analysis of interactions between membrane proteins and solutes.

New sources of cytochalasins A and B from dematiaceous hyphomycetes.

One hundred isolates of 27 species belonging to 13 genera of dematiaceous hyphomycetes were screened for production of cytochalasins A and B. Most of these isolates (94) were obtained from Assiut University Culture Collection, Botany Department, Faculty of Science, Assiut University, Egypt; three isolates from CBS, The Netherlands; two isolates from DSM, Germany; and one isolate from IMI, UK. The results revealed that 10 isolates of six species representing five genera of fungi produced cytochalasins A and/or B. These species are Alternaria chlamydospora, Cochliobolus spicifer, Diplococcum spicatum, Phoma herbarum, Phoma multipora and Setosphaeria rostrata. This is the first report for the production of cytochalasins A and/or B by these species of dematiaceous hyphomycetes.

Cytochalasin B: preparation, analysis in tissue extracts, and pharmacokinetics after intraperitoneal bolus administration in mice.

Cytochalasin B (CB) was prepared by methanol extraction of dehydrated mold (Drechslera dematioidea) matte, reverse-phase C18 silica gel batch adsorption, selective elution with 1:1 (v/v) hexane:tetrahydrofuran (THF), crystallization, preparative TLC, and recrystallization. Unit gravity silica gel normal phase chromatography afforded additional CB. Yield per liter of medium was 300 mg of CB greater than 95% pure by NMR, HPLC (60:40 hexane:THF, Lichrosorb Si60 silica gel, 230 nm), and TLC. CB added exogenously to mouse organs at 1 and 5 micrograms/organ was recovered 70 to 100% by methanol extraction, adsorption to C18 silica gel Sep-Pak cartridges, elution with ethyl acetate, and analysis by TLC and/or HPLC. Limiting sensitivity (micrograms/extract) was 0.5 TLC; 1.0 HPLC. Quantitative extraction was confirmed with 3H-labeled CB. CB ip in mice at 50 mg/kg (LD10) distributed rapidly into liver, renal fat, kidney, intestines, mesentery, pancreas, spleen, and blood cells and was cleared from all but liver within 24 h. CB was below detectable levels in thymus, lymph nodes, heart, brain, bone marrow, and lungs. Cytochalasin A is fixed to tissues and not extractable. This work affords a source of CB in quantities permitting in vivo study, provides methods for extraction and analysis, and reveals the pharmacokinetics of ip bolus CB.